Open AccessCrystallization and X‐ray diffraction studies of cellobiose phosphorylase from Cellulomonas uda
Author(s) -
Van Hoorebeke Annelies,
Stout Jan,
Kyndt John,
De Groeve Manu,
Dix Ina,
Desmet Tom,
Soetaert Wim,
Van Beeumen Jozef,
Savvides Savvas N.
Publication year - 2010
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309110002642
Subject(s) - cellobiose , disaccharide , monoclinic crystal system , crystallography , orthorhombic crystal system , chemistry , glycogen phosphorylase , glycoside hydrolase , phosphorolysis , crystallization , x ray crystallography , space group , crystal structure , stereochemistry , diffraction , enzyme , biochemistry , organic chemistry , cellulase , physics , purine nucleoside phosphorylase , purine , optics
Disaccharide phosphorylases are able to catalyze both the synthesis and the breakdown of disaccharides and have thus emerged as attractive platforms for tailor‐made sugar synthesis. Cellobiose phosphorylase from Cellulomonas uda (CP Cuda ) is an enzyme that belongs to glycoside hydrolase family 94 and catalyzes the reversible breakdown of cellobiose [β‐ d ‐glucopyranosyl‐(1,4)‐ d ‐glucopyranose] to α‐ d ‐glucose‐1‐phosphate and d ‐glucose. Crystals of ligand‐free recombinant CP Cuda and of its complexes with substrates and reaction products yielded complete X‐ray diffraction data sets to high resolution using synchrotron radiation but suffered from significant variability in diffraction quality. In at least one case an intriguing space‐group transition from a primitive monoclinic to a primitive orthorhombic lattice was observed during data collection. The structure of CP Cuda was determined by maximum‐likelihood molecular replacement, thus establishing a starting point for an investigation of the structural and mechanistic determinants of disaccharide phosphorylase activity.